PURPOSE: In this study the chondrocytic differentiation and cartilage matrix accumulation of human mesenchymal stem cells (hMSCs) were investigated after encapsulation in a genetically engineered silk-elastinlike protein polymer SELP-47 K as an injectable matrix for delivery of cell-based therapeutics. MATERIALS AND METHODS: hMSCs were encapsulated in SELP-47 K and cultured for 4 weeks in chondrogenic medium with or without transforming growth factor-beta3 (TGF). Chondrogenic differentiation was evaluated by histological, RNA and biochemical analyses for the expression of cartilage extracellular matrix components. RESULTS: Histological and immunohistochemical staining revealed that the cells acquired a rounded morphology and were embedded in significant amounts of chondrogenic extracellular matrix. Reverse transcriptase (RT)-PCR showed an up-regulation in aggrecan, type II and type X collagen and SOX9 in presence of TGF-beta3. By day 28, constructs cultured in the presence of TGF-beta3 exhibited significant increase in sulfated glycosaminoglycan and total collagen content up to 65 and 300%, respectively. CONCLUSIONS: This study demonstrates that SELP-47 K hydrogel can be used as a scaffold for encapsulation and chondrogenesis of hMSCs. The ability to use recombinant techniques to precisely control SELP structure enables the investigation of injectable protein polymer scaffolds for soft-tissue engineering with varied physicochemical properties.
PURPOSE: In this study the chondrocytic differentiation and cartilage matrix accumulation of human mesenchymal stem cells (hMSCs) were investigated after encapsulation in a genetically engineered silk-elastinlike protein polymerSELP-47 K as an injectable matrix for delivery of cell-based therapeutics. MATERIALS AND METHODS: hMSCs were encapsulated in SELP-47 K and cultured for 4 weeks in chondrogenic medium with or without transforming growth factor-beta3 (TGF). Chondrogenic differentiation was evaluated by histological, RNA and biochemical analyses for the expression of cartilage extracellular matrix components. RESULTS: Histological and immunohistochemical staining revealed that the cells acquired a rounded morphology and were embedded in significant amounts of chondrogenic extracellular matrix. Reverse transcriptase (RT)-PCR showed an up-regulation in aggrecan, type II and type X collagen and SOX9 in presence of TGF-beta3. By day 28, constructs cultured in the presence of TGF-beta3 exhibited significant increase in sulfated glycosaminoglycan and total collagen content up to 65 and 300%, respectively. CONCLUSIONS: This study demonstrates that SELP-47 K hydrogel can be used as a scaffold for encapsulation and chondrogenesis of hMSCs. The ability to use recombinant techniques to precisely control SELP structure enables the investigation of injectable protein polymer scaffolds for soft-tissue engineering with varied physicochemical properties.
Authors: J Cappello; J W Crissman; M Crissman; F A Ferrari; G Textor; O Wallis; J R Whitledge; X Zhou; D Burman; L Aukerman; E R Stedronsky Journal: J Control Release Date: 1998-04-30 Impact factor: 9.776
Authors: M F Pittenger; A M Mackay; S C Beck; R K Jaiswal; R Douglas; J D Mosca; M A Moorman; D W Simonetti; S Craig; D R Marshak Journal: Science Date: 1999-04-02 Impact factor: 47.728
Authors: Jonathan Chang; Xiu-Feng Peng; Karam Hijji; Joseph Cappello; Hamidreza Ghandehari; Santiago D Solares; Joonil Seog Journal: J Am Chem Soc Date: 2011-01-19 Impact factor: 15.419